FIGURE SUMMARY
Title

Developmental regulation of Wnt signaling by Nagk and the UDP-GlcNAc salvage pathway

Authors
Neitzel, L.R., Spencer, Z.T., Nayak, A., Cselenyi, C.S., Benchabane, H., Youngblood, C.Q., Zouaoui, A., Ng, V., Stephens, L., Hann, T., Patton, J.G., Robbins, D., Ahmed, Y., Lee, E.
Source
Full text @ Mech. Dev.
Fig. 3

Increasing levels of salvage pathway enzymes, sugars, or Dpagt1 inhibits eye formation in zebrafish. (A) (Left) Injection of DNagk or Nagk mRNAs inhibits eye formation in zebrafish. (Right) Representative images (5 days post fertilization) of disrupted eye formation in zebrafish injected with Nagk mRNA. Observed phenotypes ranged from reduced eye size to missing one or both eyes. Reduced eyes were defined as less than half the size of a wild-type eye. (B) Injection of mRNAs encoding Nagk, Pgm3, Uap1, or Dpagt1 inhibits eye formation. (C) Injection of GlcNAc, GlcNAc-6-P, GlcNAc-1-P, or UDP-GlcNAc inhibits eye formation. Results are aggregates of n ≥ 3 replicates. Injections were performed with 0.5 ng mRNA or 25 pmol sugar. Significance was calculated using Fisher's exact test with Bonferroni correction. Absolute numbers are indicated above bars. (A) *ρ < 0.0253 and **ρ < 0.00501. (B–C) *ρ < 0.0127, **ρ < 0.00251, and *** ρ < 0.000251.

 
Fig. 4

Inhibition of Nagk and N-glycosylation in zebrafishinduces cyclocephaly. (A) (Left) Injection of NagkT128M mRNA or 3-OMe-GlcNAc results in cyclopic zebrafish. (Right) Representative images of cyclopic zebrafish (5 days post fertilization) injected with NagkT128M mRNA. Animals were defined as cyclopic as previously described (Marlow et al., 1998). *ρ < 0.0253. (B) Injection of NagkT128M mRNA induces cyclopia, which is rescued by co-injection with GlcNAc, GlcNAc-6-P, GlcNAc-1-P, or UDP-GlcNAc. *ρ < 0.0127. (C) Injection of Ngly1mRNA also induces cyclopia. *ρ < 0.05. Results are aggregates of n ≥ 3 replicates. Injections were performed with 0.5 ng mRNA or 25 pmol sugar. Absolute numbers are indicated above bars. Significance was calculated using Fisher's exact test with Bonferroni correction.

 
Acknowledgments
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Reprinted from Mechanisms of Development, 156, Neitzel, L.R., Spencer, Z.T., Nayak, A., Cselenyi, C.S., Benchabane, H., Youngblood, C.Q., Zouaoui, A., Ng, V., Stephens, L., Hann, T., Patton, J.G., Robbins, D., Ahmed, Y., Lee, E., Developmental regulation of Wnt signaling by Nagk and the UDP-GlcNAc salvage pathway, 20-31, Copyright (2019) with permission from Elsevier. Full text @ Mech. Dev.
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